S-acylated Golga7b stabilises DHHC5 at the plasma membrane to regulate desmosome assembly and cell adhesion

Woodley, Keith; Collins, Mark O.

doi:10.1101/481861

Cited by 19 publications

(50 citation statements)

References 46 publications

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“…Several recent studies have shown that the C-terminal S -acylated cysteines in zDHHC5 ( Fig. 2 ) are important in regulating the distribution of this enzyme between the plasma membrane and internal membranes in nonneuronal cells ( 85 – 87 ). Indeed, one study reported that S -acylation of these cysteines occurred in response to isoproterenol stimulation, leading to stabilization of zDHHC5 at the plasma membrane ( 85 ).…”

Section: Regulation Of Zdhhc Enzymes By S -Acylatimentioning

confidence: 99%

Regulatory effects of post-translational modifications on zDHHC S-acyltransferases

Zmuda

Chamberlain

2020

Journal of Biological Chemistry

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The human zDHHC S-acyltransferase family comprises 23 enzymes that mediate the S-acylation of a multitude of cellular proteins, including channels, receptors, transporters, signalling molecules, scaffolds and chaperones. This reversible post-transitional modification (PTM) involves the attachment of a fatty acyl chain, usually derived from palmitoyl-CoA, to specific cysteine residues on target proteins, which affects their stability, localisation and function. These outcomes are essential to control many processes, including synaptic transmission and plasticity, cell growth and differentiation, and infectivity of viruses and other pathogens. Given the physiological importance of S-acylation, it is unsurprising that perturbations in this process, including mutations in ZDHHC genes, have been linked to different neurological pathologies and cancers, and there is growing interest in zDHHC enzymes as novel drug targets. Although zDHHC enzymes control a diverse array of cellular processes and are associated with major disorders, our understanding of these enzymes is surprisingly incomplete, particularly in regards to the regulatory mechanisms controlling these enzymes. However, there is growing evidence highlighting the role of different PTMs in this process. In this review, we discuss how PTMs including phosphorylation, S-acylation and ubiquitination affect the stability, localisation and function of zDHHC enzymes and speculate on possible effects of PTMs that have emerged from larger screening studies. Developing a better understanding of the regulatory effects of PTMs on zDHHC enzymes will provide new insight into the intracellular dynamics of S-acylation and may also highlight novel approaches to modulate S-acylation for clinical gain.

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Section: Regulation Of Zdhhc Enzymes By S -Acylatimentioning

confidence: 99%

Regulatory effects of post-translational modifications on zDHHC S-acyltransferases

Zmuda

Chamberlain

2020

Journal of Biological Chemistry

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“…Golga7b is palmitoylated by zDHHC5 and interacts with the same region of the zDHHC5 C tail as the Na-pump in a palmitoylation-dependent manner. Palmitoylated Golga7b is required for delivery of zDHHC5 to the cell surface as well as recruitment to and palmitoylation of substrates by zDHHC5 23 . When we carried out a detailed proteomic analysis of the proteins in cardiac muscle that interact with NBSP, however, neither Golga7 nor Golga7b were identified 48 .…”

Section: Discussionmentioning

confidence: 99%

“…However, this does not rule out the possibility of zDHHC5 and the Na-pump α-subunit interacting indirectly via an adapter protein. Recently the activity and localization of zDHHC5 in HEK and HeLa cells has been found to be regulated by two closely related proteins, Golga7 47 and Golga7b 23 respectively. Golga7b is palmitoylated by zDHHC5 and interacts with the same region of the zDHHC5 C tail as the Na-pump in a palmitoylation-dependent manner.…”

Section: Discussionmentioning

confidence: 99%

“…The acyltransferase zDHHC5 is primarily located at the cell surface 19 where it regulates a diverse range of physiological processes. Known zDHHC5 substrates include somatostatin receptor 5 20 , sphingosine-1-phosphate receptor 1 21 , the lipid raft protein flotillin 2 22 , the Na-pump regulatory protein phospholemman (PLM) 7,13 , desmoglein-2 and plakophilin-3 23 as well as the neuronal proteins δ-catenin 24 and GRIP 1 25 .…”

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confidence: 99%

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Control of protein palmitoylation by regulating substrate recruitment to a zDHHC-protein acyltransferase

et al. 2020

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Although palmitoylation regulates numerous cellular processes, as yet efforts to manipulate this post-translational modification for therapeutic gain have proved unsuccessful. The Napump accessory sub-unit phospholemman (PLM) is palmitoylated by zDHHC5. Here, we show that PLM palmitoylation is facilitated by recruitment of the Na-pump α sub-unit to a specific site on zDHHC5 that contains a juxtamembrane amphipathic helix. Site-specific palmitoylation and GlcNAcylation of this helix increased binding between the Na-pump and zDHHC5, promoting PLM palmitoylation. In contrast, disruption of the zDHHC5-Na-pump interaction with a cell penetrating peptide reduced PLM palmitoylation. Our results suggest that by manipulating the recruitment of specific substrates to particular zDHHC-palmitoyl acyl transferases, the palmitoylation status of individual proteins can be selectively altered, thus opening the door to the development of molecular modulators of protein palmitoylation for the treatment of disease.

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“…LC-MS/MS analysis was performed and data was processed and quantified as described previously 62 . Briefly, peptides were analysed by nanoflow LC-MS/MS using an Orbitrap Elite (Thermo Fisher) hybrid mass spectrometer equipped with a nanospray source, coupled to an Ultimate RSLCnano LC System (Dionex).…”

Section: Methodsmentioning

confidence: 99%

DLG2 knockout reveals neurogenic transcriptional programs underlying neuropsychiatric disorders and cognition

Sanders

D’Andrea

Collins³

et al. 2020

Preprint

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Genetic studies robustly implicate perturbation of DLG2-scaffolded mature postsynaptic signalling complexes in schizophrenia. Here we study in vitro cortical differentiation of DLG2 -/human embryonic stem cells via integrated phenotypic, gene expression and disease genetic analyses. This uncovers a developmental role for DLG2 in the regulation of neural stem cell proliferation and adhesion, and the activation of transcriptional programs during early excitatory corticoneurogenesis. Down-regulation of these programs in DLG2 -/lines delays expression of cell-type identity and causes marked deficits in neuronal migration, morphology and active properties. Genetic risk factors for neuropsychiatric and neurodevelopmental disorders converge on these neurogenic programs, each disorder displaying a distinct pattern of enrichment. These data unveil an intimate link between neurodevelopmental and mature signalling deficits contributing to disease -suggesting a dual role for known synaptic risk genes -and reveal a common pathophysiological framework for studying the neurodevelopmental origins of Mendelian and genetically complex mental disorders. Results Knockout generation and validationTwo DLG2 -/lines were created from H7 hESCs using the CRISPR/Cas9-D10A nickase system targeting the first PDZ domain ( Figure S1). Sequencing revealed no off-target mutations (see Methods, Figure S2 & Table S1). A significant decrease in DLG2 mRNA was observed for exons spanning the first PDZ domain, with a similar decrease inferred for PDZ-containing transcripts, indicating degradation of DLG2 -/transcripts via nonsense-mediated decay ( Figure S3A, B). Quantitative mass spectrometry-based proteomic analysis of peptide-affinity pulldowns using the NMDA receptor NR2 subunit PDZ peptide ligand (Husi and Grant, 2001) confirmed the presence of DLG2 in pulldowns from WT but not DLG2 -/lines ( Figure S3C-F & Table S2). Genotyping revealed no CNVs in either DLG2 -/line relative to WT ( Figure S4A). Both DLG2 -/lines expressed pluripotency markers OCT4, SOX2 and NANOG at 100% of WT levels ( Figure S4B-E). Cells were extensively characterised for their cortical identity using western blotting and immunocytochemistry from day 20 to 60 ( Figure S5). Over 90% of day 20 cells were positive for FOXG1, PAX6 and SOX2 confirming their dorsal forebrain fate ( Figure S5A-B). DLG2 regulates neural stem-cell proliferation and adhesionRNA was extracted in triplicate from each line at 4 timepoints spanning cortical excitatory neuron development (Figure 1A, B) and gene expression quantified. To robustly identify genes dysregulated by DLG2 knockout, expression data from the 2 DLG2 -/lines were pooled and compared to a WT sister line at each timepoint (see Methods). Disruption of DLG2 had a profound effect: of the >13,000 protein-coding genes expressed at each timepoint, ~7% were differentially expressed between DLG2 -/and WT at day 15, rising to 40-60% between days 20 and 30 then decreasing to ~25% by day 60 ( Figure 1C). Strikingly, the 3 genes with the strongest ...

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S-acylated Golga7b stabilises DHHC5 at the plasma membrane to regulate desmosome assembly and cell adhesion

Cited by 19 publications

References 46 publications

Regulatory effects of post-translational modifications on zDHHC S-acyltransferases

Regulatory effects of post-translational modifications on zDHHC S-acyltransferases

Control of protein palmitoylation by regulating substrate recruitment to a zDHHC-protein acyltransferase

DLG2 knockout reveals neurogenic transcriptional programs underlying neuropsychiatric disorders and cognition

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